What is your diagnosis. Bone sequestrum of the proximoplantarolateral aspect of the tuber calcaneus.
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Biomedical subjects
M M Moon
Publications and source records attributed to M M Moon.
Analysis of adhesion, piliation, protease production and ocular infectivity of several P. aeruginosa strains.
The role of bacterial piliation and protease production in Pseudomonas aeruginosa adhesion to the injured corneal epithelial surface and subsequent infectivity was examined using several bacterial strains, including three that were hyperpiliated. To initiate this study, bacteria were examined by transmission EM to confirm their piliation characteristics. The PAK strain, like pseudomonas ATCC 19660, possessed about 1-4 polar pili. The mutant PAK/PR11 lacked pili while PAK/PR1, DB2, a mutant of PAO1, and PA1244, a wild-type clinical isolate, were hyperpiliated. Ocular infectivity of these bacterial strains and mutants was examined macroscopically and histopathologically in mice and these data compared to the well-characterized ocular disease response of a murine model of infection with pseudomonas ATCC 19660. The PAK strain was infective, but less virulent than strain 19660 by both macroscopic grading and histopathological analysis of infected eyes. Infectivity of the PR11 mutant was similar to the PAK parent strain, while PR1, DB2 and 1244, all hyperpiliated, were not infective. To explore the hypothesis that hyperpiliated bacteria bound less well to cornea and thus failed to induce corneal disease, in vitro quantitative studies of bacterial adhesion were done using an ocular organ culture model. The PR1 hyperpiliated mutant bound significantly less well to cornea than the PAK parent strain, PR11 mutant or pseudomonas 19660, while DB2 and 1244 binding did not differ significantly from 19660 or PAK. Examination of protease production, another factor which may influence adhesion, revealed that only 19660 and DB2 produced detectable protease. This study provides evidence that non-piliated, non-protease producing strains such as PAK/PR11 possess alternate virulence mechanisms to facilitate binding to and infectivity of corneal tissue.
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Monoclonal antibodies provide protection against ocular Pseudomonas aeruginosa infection.
A panel of well characterized monoclonal antibodies (MAbs) directed against outer membrane proteins H2, or F (porin) of Pseudomonas aeruginosa were examined to determine whether they exhibited any protective effect against subsequent ocular challenge with the bacteria topically applied to the scarified corneal surface. Mice were observed macroscopically following bacterial challenge and the degree of ocular disease graded on a scale of 0 to 4 (0, normal, fully protected cornea; 4, corneal perforation or phthisis, not protected). Mice treated intravenously with either MAb MA1-6 (anti-H2) or MA2-10 (anti-F), or a combination of these two MAbs and MAb MA4-4 (anti-F), two hours before corneal challenge with the viable bacteria, exhibited significantly less corneal disease than mice either not treated with the MAbs, treated with MA4-4 alone or treated with MAb MA1-3 (anti-I). The latter MAb is directed against an outer membrane epitope that is not surface exposed. Light and transmission electron microscopic histopathology also was employed and provided confirmatory evidence to support the macroscopic analyses.
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Ocular surface complex carbohydrates are modified with aging.
Picric acid-paraformaldehyde-glutaraldehyde (PA-P-G) was used to stabilize chemically ocular surface-associated mucus in mice of various ages. Transmission electron microscopy was then used to examine those specimens stained with cationic ferritin (CF), dialysed iron and Alcian Blue. Collectively, all of these stains are markers for anionic sulfate or carboxyl groups. With each of them, positive labeling of the ocular surface was observed for all ages examined, even when mucus cannot be morphologically demonstrated. Except for dialysed iron, staining was weak in the youngest animals and heaviest in young adult and aged mice. The ocular surface was negative for high iron diamine (HID) in pups and older mice through 1 year of age. Scant positive staining for HID was seen at the ocular surface in 14-month-old mice indicating that mucus became slightly sulfated with aging. Treatment of eyes with neuraminidase prior to fixation reduced the number of CF binding sites in all ages of mice examined, confirming that many of the carboxyl groups at the ocular surface are associated with sialic acid residues. Comparison of percentage reduction in CF labeling following neuraminidase treatment of the eyes of 5- and 10-postnatal day mice with all other ages of mice suggested that fewer removable carboxyl groups at the ocular surface are associated with sialic acid residues in pups. The ocular surface of all eyes also stained positively at the TEM level when a periodic acid-thiocarbohydrazide-silver protein (PA-T-SP) staining sequence was used. Collectively, these data are of significance with respect to further characterization of the ocular surface, particularly with regard to development-associated changes and their possible role in defence of the eye surface.
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Evidence for N-acetylmannosamine as an ocular receptor for P. aeruginosa adherence to scarified cornea.
This study provides evidence for an N-acetylmannosamine (manNac) receptor mediating in vivo bacterial adherence of P. aeruginosa to the scarified corneal epithelial surface. Bacteria, whether exposed to the inhibiting sugar or not, did not adhere immediately after inoculation, but required time in contact with the scarified corneal surface to adhere and adherence increased with time. Organisms were observed primarily adherent to the surface of epithelial cells lining the wound and were less frequently seen on areas of denuded stroma. Saturation of binding sites on the bacterial surface by manNac inhibited attachment of the organisms to newly exposed surface membrane receptors. In vivo protection studies showed excellent correlation with quantitative analysis of scanning electron micrographs, in that the number of adherent organisms at the corneal surface at 60 min following scarification and bacterial inoculation was decreased significantly by prior treatment of the bacteria with manNac when compared with PBS or the other monosaccharides. These data were confirmed by quantitative bacterial culture of excised corneas which had been similarly infected in vivo with PBS or manNac treated organisms.
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Age alters ADPase positive dendritic (Langerhans) cell response to P. aeruginosa ocular challenge.
The morphology, distribution and quantitation of dendritic (Langerhans) cells (LC) was determined by analysis of ADPase stained epithelial flat mounts from 6-8 week young adult (resistant) and 24 month old (susceptible) aged mice before and after experimental infection with P. aeruginosa topically applied to the scarified cornea. The contralateral eye (controls) was also scarified and phosphate buffered saline applied similarly. This study has examined the changes in ADPase positive cell populations of the conjunctival limbal epithelium and corneal epithelium of naturally resistant mice (Swiss-Webster and CD2F1) following corneal infection with Pseudomonas aeruginosa at two different ages, young adult (8 week old) and aged (24 month old). The young adult mice recover from their infection and restore corneal clarity while the aged mice have extensive ocular destruction and corneal scarring. Conjunctival limbal dendritic cell numbers in young adult mice were found to be significantly increased at day seven post infection and then returned to baseline levels. In contrast, conjunctival limbal dendritic cell numbers in aged mice were found to increase slowly and to peak at fourteen days after infection. Other differences between the two ages (young adult and aged) included an initial increase in dendritic cells five hours post infection in the young adult groups and an initial decrease at five hours in the aged groups of mice.
Aging↗
Requirements for inducing tolerance of hapten-specific delayed hypersensitivity: epitope density.
Tolerance to hapten-specific antibody formation and delayed hypersensitivity was examined in adult rabbits. The azobenzenearsonate (ABA) or sulfonate-specific antibody response to hapten-hemocyanin immunogens was suppressed by deaggregated hapten-rabbit IgG conjugates given 21 and 14 days before challenge. High affinity antibody was preferentially suppressed. Delayed hypersensitivity to ABA-tyrosine was suppressed by deaggregated ABA-rabbit IgG conjugates injected 17 and 10 days before challenge. Conjugates with a high hapten density, ABA15-23-rabbit IgG were effective tolerogens. Conjugates with four to six ABA groups per carrier molecule were very poor tolerogens. Increasing the amount of low substituted conjugate injected did not improve tolerogenicity. It appears that a high epitope density is required for effective induction of tolerance to ABA-specific delayed hypersensitivity in the rabbit.
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